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Submitted to: Biofuels
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 4/14/2010 Publication Date: 7/1/2010 Citation: Lee, C.C. 2010. Screening assays for biomass-degrading enzymes. Biofuels. 1(4):575-588. Interpretive Summary: Biomass is a valuable resource for renewable products, such as fuel. In order to harness this material, the biomass must be broken down with a vast array of distinct enzymes. This manuscript describes the various screening strategies that can be used to isolate enzymes that hydrolyze the cellulose and hemicellulose (specifically xylan) fractions of biomass. These assays can also be used in directed evolution projects to engineer improved enzyme characteristics. Technical Abstract: The enzymes that break down the components of lignocellulosic biomass play an important role in the production of value-added chemical feedstocks and biofuels. In order to accommodate the variety of substrates and industrial process conditions, enzymes with diverse activity profiles are required. These enzymes are encoded by genes obtained either through screening of microorganisms or genetic engineering of cloned genes by techniques such as directed evolution. Both of these strategies require activity assays that facilitate the screening of many recombinants to isolate those with the desired characteristics. In this manuscript, activity assays for enzymes that degrade cellulose and hemicellulose (specifically xylan) are described for analyzing samples at various throughputs. |
